Purification and multimer formation of allurin, a sperm chemoattractant from Xenopus laevis egg jelly.
Allurin, a sperm chemoattractant isolated from Xenopus laevis egg jelly, can be purified in one step from an extract of diffusible jelly proteins ("egg water") using a FPLC or HPLC anion exchange column and a multi-step NaCl gradient. Allurin homomultimers were detected by Western blotting with antibodies prepared against the purified protein or peptides within the protein. Allurin multimers were stable and resisted dissociation by SDS and beta-mercaptoethanol. Alkylation of allurin provided evidence for two free sulfhydryl groups but did not eliminate multimer formation, suggesting that intermolecular disulfide bond formation is not required for allurin aggregation. Concentration of egg water was accompanied by a reduction of chemoattractant activity that could not be fully accounted for by homomultimer formation. Rather, the presence of a multiphasic dose-activity curve upon partial purification and formation of hetero-allurin complexes during concentration suggested that egg water may contain allurin-binding proteins that reduce multimer formation and activity.
Sugiyama
Hitoshi
H
Department of Physiology, St. Marianna University School of Medicine, Kawasaki, Japan.
Burnett
Lindsey
L
Xiang
Xueyu
X
Olson
John
J
Willis
Shaun
S
Miao
Amy
A
Akema
Tatsuo
T
Bieber
Allan L
AL
Chandler
Douglas E
DE
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D002630:Chemotactic Factors / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D002633:Chemotaxis / Q000502:physiology; D004527:Egg Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D005260:Female; D008297:Male; D009865:Oocytes / Q000737:chemistry*; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013094:Spermatozoa / Q000378:metabolism*; D014982:Xenopus laevis
D002352:Carrier Proteins; D002630:Chemotactic Factors; D004527:Egg Proteins; C438377:allurin